激光微照射后蛋白质迁移和定位的高通量评估。

Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT, 06520, USA.

Institute of Cell Biology (Cancer Research), University Hospital Essen, University of Duisburg-Essen, Essen, 45122, Germany.

Sci Rep. 2019 Feb 28;9(1):3148. doi: 10.1038/s41598-019-39760-8.

DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn't accumulate in foci, laser micro-irradiation of single nuclei provides an alternative method to monitor DNA repair proteins and histone dynamics at the DNA damage site. We have developed an automated evaluation tool for standardized, high-throughput analysis of micro-irradiated cells featuring single cell background subtraction and detection across multiple fluorescence channels, allowing for robust statistics.

DNA 和组蛋白相关的研究通常包括对蛋白质修饰（如组蛋白磷酸化）的定量分析。分析蛋白质焦点的积累和消失可用于监测 DNA 损伤和修复动力学。如果感兴趣的蛋白质没有在焦点中积累，那么激光微照射单个核就为监测 DNA 修复蛋白和组蛋白在 DNA 损伤部位的动力学提供了一种替代方法。我们开发了一种自动化评估工具，用于对微照射细胞进行标准化、高通量分析，具有单细胞背景扣除和多个荧光通道的检测功能，可实现强大的统计学分析。